Lee J W, Kim K m, Jung S H, Lee K J, Choi E C, Sung Y C, Kang C Y
Laboratory of Immunology, College of Pharmacy, Seoul National University, Seoul 151-742, Korea.
J Virol. 1999 Jan;73(1):11-8. doi: 10.1128/JVI.73.1.11-18.1999.
Evidence from clinical and experimental studies of human and chimpanzees suggests that hepatitis C virus (HCV) envelope glycoprotein E2 is a key antigen for developing a vaccine against HCV infection. To identify B-cell epitopes in HCV E2, six murine monoclonal antibodies (MAbs), CET-1 to -6, specific for HCV E2 protein were generated by using recombinant proteins containing E2t (a C-terminally truncated domain of HCV E2 [amino acids 386 to 693] fused to human growth hormone and glycoprotein D). We tested whether HCV-infected sera were able to inhibit the binding of CET MAbs to the former fusion protein. Inhibitory activity was observed in most sera tested, which indicated that CET-1 to -6 were similar to anti-E2 antibodies in human sera with respect to the epitope specificity. The spacial relationship of epitopes on E2 recognized by CET MAbs was determined by surface plasmon resonance analysis and competitive enzyme-linked immunosorbent assay. The data indicated that three overlapping epitopes were recognized by CET-1 to -6. For mapping the epitopes recognized by CET MAbs, we analyzed the reactivities of CET MAbs to six truncated forms and two chimeric forms of recombinant E2 proteins. The data suggest that the epitopes recognized by CET-1 to -6 are located in a small domain of E2 spanning amino acid residues 528 to 546.
来自人类和黑猩猩的临床及实验研究证据表明，丙型肝炎病毒（HCV）包膜糖蛋白E2是开发抗HCV感染疫苗的关键抗原。为了鉴定HCV E2中的B细胞表位，通过使用含有E2t（HCV E2的C末端截短结构域[氨基酸386至693]与人生长激素和糖蛋白D融合）的重组蛋白，产生了六种针对HCV E2蛋白的鼠单克隆抗体（MAb），即CET-1至-6。我们测试了HCV感染血清是否能够抑制CET MAb与前一种融合蛋白的结合。在大多数测试血清中观察到抑制活性，这表明就表位特异性而言，CET-1至-6与人类血清中的抗E2抗体相似。通过表面等离子体共振分析和竞争性酶联免疫吸附测定法确定了CET MAb识别的E2上的表位的空间关系。数据表明，CET-1至-6识别三个重叠表位。为了绘制CET MAb识别的表位图谱，我们分析了CET MAb对重组E2蛋白的六种截短形式和两种嵌合形式的反应性。数据表明，CET-1至-6识别的表位位于E2的一个小结构域中，该结构域跨越氨基酸残基528至546。
